Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-AMF antibody [EPR11663(B)] (ab167394)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR11663(B)] to AMF
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-AMF antibody [EPR11663(B)]
    See all AMF primary antibodies

  • Description

    Rabbit monoclonal [EPR11663(B)] to AMF

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide corresponding to Human AMF.
    Database link: P06744

  • Positive control

    • WB: HEK-293T, HepG2, HeLa, HT29, and A549 cell lysates. IHC-P: Human brain, pancreas, skeletal muscle, ovarian carcinoma, prostatic hyperplasia and thyroid carcinoma tissues. ICC/IF: MCF7 and HepG2 cells. Flow Cyt: A549 cells.

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

     This product was previously labelled as Glucose 6 phosphate isomerase

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab167394 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/10000. Predicted molecular weight: 63 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/500.
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
  • Application notes
    Is unsuitable for IP.

    • Target

    • Function

      Besides it's role as a glycolytic enzyme, mammalian GPI can function as a tumor-secreted cytokine and an angiogenic factor (AMF) that stimulates endothelial cell motility. GPI is also a neurotrophic factor (Neuroleukin) for spinal and sensory neurons.

    • Pathway

      Carbohydrate degradation; glycolysis; D-glyceraldehyde 3-phosphate and glycerone phosphate from D-glucose: step 2/4.

    • Involvement in disease

      Defects in GPI are the cause of hemolytic anemia non-spherocytic due to glucose phosphate isomerase deficiency (HA-GPID) [MIM:613470]. It is a form of anemia in which there is no abnormal hemoglobin or spherocytosis. It is caused by glucose phosphate isomerase deficiency. Severe GPI deficiency can be associated with hydrops fetalis, immediate neonatal death and neurological impairment.

    • Sequence similarities

      Belongs to the GPI family.

    • Post-translational
      modifications

      Phosphorylation at Ser-185 by CK2 has been shown to decrease enzymatic activity and may contribute to secretion by a non-classical secretory pathway.
      ISGylated.

    • Cellular localization

      Cytoplasm. Secreted.
    • Information by UniProt

    • Database links

      • Alternative names

        • AMF antibody
        • Aurocrine motility factor antibody
        • Autocrine motility factor antibody
        • DKFZp686C13233 antibody
        • EC 5.3.1.9 antibody
        • G6PI_HUMAN antibody
        • Glucose phosphate isomerase antibody
        • Glucose-6-phosphate isomerase antibody
        • GNPI antibody
        • GPI antibody
        • Gpi1 antibody
        • Hexose monophosphate isomerase antibody
        • Hexosephosphate isomerase antibody
        • Neuroleukin antibody
        • NLK antibody
        • Oxoisomerase antibody
        • PGI antibody
        • PHI antibody
        • Phosphoglucose isomerase antibody
        • Phosphohexomutase antibody
        • Phosphohexose isomerase antibody
        • Phosphosaccharomutase antibody
        • SA 36 antibody
        • SA-36 antibody
        • SA36 antibody
        • Sperm antigen 36 antibody
        see all

      Images

      • Western blot - Anti-AMF antibody [EPR11663(B)] (ab167394)
        Western blot - Anti-AMF antibody [EPR11663(B)] (ab167394)
        All lanes : Anti-AMF antibody [EPR11663(B)] (ab167394) at 1/1000 dilution

        Lane 1 : Wild-type HEK293T cell lysate
        Lane 2 : GPI knockout HEK293T cell lysate
        Lane 3 : HepG2 cell lysate
        Lane 4 : HeLa cell lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

        Predicted band size: 63 kDa
        Observed band size: 63 kDa



        Lanes 1-4: Merged signal (red and green). Green - ab167394 observed at 63 kDa. Red - loading control ab8245 observed at 36 kDa.

         ab167394 Anti-AMF antibody [EPR11663(B)] was shown to specifically react with AMF in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266834 (knockout cell lysate ab257458) was used. Wild-type and AMF knockout samples were subjected to SDS-PAGE. ab167394 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

      • Immunocytochemistry/ Immunofluorescence - Anti-AMF antibody [EPR11663(B)] (ab167394)
        Immunocytochemistry/ Immunofluorescence - Anti-AMF antibody [EPR11663(B)] (ab167394)

        Immunocytochemistry/Immunofluorescence analysis of MCF-7 (human breast carcinoma) labelling AFM with purified ab167394 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

        Control: PBS only

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)

        ab167394 showing +ve staining in Human normal brain.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Flow Cytometry - Anti-AMF antibody [EPR11663(B)] (ab167394)
        Flow Cytometry - Anti-AMF antibody [EPR11663(B)] (ab167394)

        Flow cytometric analysis of permeabilized A549 cells, labeling AFM with ab167394 at 1/10 dilution.

      • Western blot - Anti-AMF antibody [EPR11663(B)] (ab167394)
        Western blot - Anti-AMF antibody [EPR11663(B)] (ab167394)
        All lanes : Anti-AMF antibody [EPR11663(B)] (ab167394) at 1/1000 dilution

        Lane 1 : HepG2 cell lysate
        Lane 2 : HeLa cell lysate
        Lane 3 : HT29 cell lysate
        Lane 4 : A549 cell lysate

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : Goat anti-rabbit HRP at 1/2000 dilution

        Predicted band size: 63 kDa

      • Immunocytochemistry/ Immunofluorescence - Anti-AMF antibody [EPR11663(B)] (ab167394)
        Immunocytochemistry/ Immunofluorescence - Anti-AMF antibody [EPR11663(B)] (ab167394)

        Immunofluorescent analysis of HepG2 cells labeling AFM with ab167394 at 1/100 dilution.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)

        ab167394 showing +ve staining in Human normal pancreas.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)

        ab167394 showing +ve staining in Human ovarian carcinoma.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)

        ab167394 showing +ve staining in Human skeletal muscle.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)

        Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue, labeling AFM with ab167394 at 1/100 dilution.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)

        Immunohistochemical analysis of paraffin-embedded Human thyroid carcinoma tissue labeling AFM with ab167394 at 1/100 dilution.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Anti-AMF antibody [EPR11663(B)] (ab167394)
        Anti-AMF antibody [EPR11663(B)] (ab167394)

      References (1)

      Publishing research using ab167394? Please let us know so that we can cite the reference in this datasheet.

      ab167394 has been referenced in 1 publication.

      • He Q  et al. Mesenchymal stem cell-derived exosomes exert ameliorative effects in type 2 diabetes by improving hepatic glucose and lipid metabolism via enhancing autophagy. Stem Cell Res Ther 11:223 (2020). PubMed: 32513303

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